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使用新型抗菌剂重组人β-防御素-3对颌面硅橡胶弹性体进行消毒

Disinfection of maxillofacial silicone elastomer using a novel antimicrobial agent: recombinant human beta-defensin-3.

作者信息

Shi Y, Song W, Feng Z H, Zhao Y T, Li F, Tian Y, Zhao Y M

机构信息

Department of Prosthodontics, Fourth Military Medical University, Xi'an, Shaanxi Province, China.

出版信息

Eur J Clin Microbiol Infect Dis. 2009 Apr;28(4):415-20. doi: 10.1007/s10096-008-0634-z. Epub 2008 Oct 8.

Abstract

Maxillofacial silicone elastomer, when used as a prosthesis, is in contact with wound surfaces and mucosa, and tends to be contaminated with microorganisms from a patient's saliva and blood. The aim of the study was to evaluate the efficacy of human beta-defensin-3 (HBD3) on the reduction of two resistant bacteria species from the surface of maxillofacial silicone elastomer. HBD3 cDNA was amplified from total RNA, which had been extracted from human gingival epithelium by means of reverse-transcription polymerase chain reaction (RT-PCR). Following this, the cDNA fragments were recombined in a prokaryotic expression vector. The constructed expression vectors pET-32a/HBD3 were transformed into Escherichia coli to obtain recombinant protein. After protein purification and refolding, the product was verified in classic antimicrobial experiments against Staphylococcus aureus and Candida albicans. Specimens made of silicone elastomer A-2186, which had been contaminated with S. aureus or C. albicans, were immersed in rHBD3 or 5.25% sodium hypochlorite (a positive control) for 5 min, 10 min, 30 min, or 60 min. The active recombinant HBD3 obtained in the current study eliminated the S. aureus and C. albicans microorganism from the surface of the maxillofacial elastomer after a 30-min immersion. There was no statistically significant difference between the rHBD3 group and the sodium hypochlorite 5.25% group. In conclusion, rHBD3 exhibits antibacterial activity against oral pathogenic strains that adhere to maxillofacial elastomer, and may, thus, contribute to the prevention of infections caused by S. aureus and C. albicans.

摘要

颌面硅橡胶弹性体用作假体时,会与伤口表面和黏膜接触,容易被患者唾液和血液中的微生物污染。本研究的目的是评估人β-防御素3(HBD3)对减少颌面硅橡胶弹性体表面两种耐药菌的效果。通过逆转录聚合酶链反应(RT-PCR)从人牙龈上皮提取的总RNA中扩增HBD3 cDNA。随后,将cDNA片段重组到原核表达载体中。将构建的表达载体pET-32a/HBD3转化到大肠杆菌中以获得重组蛋白。经过蛋白纯化和复性后,在针对金黄色葡萄球菌和白色念珠菌的经典抗菌实验中对产物进行验证。将被金黄色葡萄球菌或白色念珠菌污染的由硅橡胶弹性体A-2186制成的标本浸入重组人HBD3或5.25%次氯酸钠(阳性对照)中5分钟、10分钟、30分钟或60分钟。本研究中获得的活性重组HBD3在浸泡30分钟后消除了颌面弹性体表面的金黄色葡萄球菌和白色念珠菌微生物。重组人HBD3组与5.25%次氯酸钠组之间无统计学显著差异。总之,重组人HBD3对粘附于颌面弹性体的口腔致病菌株具有抗菌活性,因此可能有助于预防由金黄色葡萄球菌和白色念珠菌引起的感染。

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